An Online mtDNA Tool for Identification of Neotropical Psittacid Species and Taxonomic Issues:A Study Case of the Amazona ochrocephala Complex

Parrots are among the most popular pets in the world and they are also some of the most illegally traded, particularly in Brazil. Some computational tools were recently developed by researchers based on molecular databases for taxonomy support, forensic identification and conservation purposes. In this study, the DNA Surveillance platform was used to build an online database tool for molecular identification of Brazilian Psittacids using DNA sequences of six mitochondrial genes. To illustrate possible taxonomic issues of the online tool due to interspecific hybridization or unresolved taxonomy, we focused on Amazona aestiva that is considered as one of the most common parrots in Brazil, commonly bred as pets, and considered to be part of a species complex with Amazona ochrocephala from South America. We provide three curated sequence databases, which allow the species identification of individuals or tissue samples of birds of the Psittacidae family using mitochondrial DNA markers, and a comprehensive description of a taxonomic issue involving the A. ochrocephala complex. The results obtained corroborate previous studies suggesting that these species are not reciprocally monophyletic, due to either an ancient hybridization in central Brazil, or, they maybe just are morpho-varieties of the same species. Alternatively, if A. aestiva and A. ochrocephala were considered as sister species, the data could be interpreted either as a result of secondary contact or incipient speciation. Beyond the use of mtDNA for species identification, the high mtDNA haplotype diversity observed in A. aestiva indicates its potential use in discrimination of lineages that could be an important auxiliary tool to certify the captive origin of legally commercialized parrots.

Impact of hormonal modulation at proestrus on ovarian responses and uterine gene expression of suckled anestrous beef cows

Background： This study evaluated the impact of hormonal modulation at the onset of proestrus on ovarian response and uterine gene expression of beef cows.Methods： A total of 172 anestrous beef cows were assigned to one of four groups according to the treatment with estradiol cypionate（ECP） and/or equine chorionic gonadotropin（eCG） [CON（n = 43）, ECP（n = 43）, eCG（n = 44）and ECP ＋ eCG（n = 42）].Results： ECP-treated cows（ECP and ECP ＋ eCG groups） presented greater occurrence of estrus（44.6% vs. 65.4%; P = 0.01） and pregnancy per AI [47.1% vs. 33.3%; P = 0.07], but similar progesterone（P4） concentration at subsequent diestrus than cows not treated with ECP（CON and eCG groups）. Nonetheless, eCG-treated cows（eCG and ECP ＋ eCG groups） presented larger follicle at timed AI（12.6 ± 0.3 vs. 13.5 ± 0.3 mm; P = 0.03）, greater ovulation rate（96.5%vs. 82.6%; P = 0.008） and greater P4 concentration at d 6（3.9 ± 0.2 vs. 4.8 ± 0.2 ng/mL; P = 0.001） than cows not treated with eCG（CON and ECP groups）. Next, cows with a new corpus luteum 6 d after TAI were submitted to uterine biopsy procedure. Uterine fragments [CON（n = 6）, ECP（n = 6）] were analyzed by RNA-Seq and a total of 135 transcripts were differentially expressed between groups（73 genes up-regulated by ECP treatment）. Subsequently,uterine samples were analyzed by q PCR（genes associated with cell proliferation）. ECP treatment induced greater abundance of PTCH2（P = 0.07） and COL4 A1（P = 0.02）, whereas suppressed EGFR（P = 0.09） expression. Conversely, eCG treatment increased abundance of HB-EGF（P = 0.06）, ESR2（P = 0.09）, and ITGB3（P = 0.05）, whereas it reduced transcription of ESR1（P = 0.05）. Collectively, supplementation with ECP or eCG at the onset of proestrous of anestrous beef cows influenced ovarian responses, global and specific endometrial gene expression.Conclusion： Proestrus estradiol regulate the endometrial transcriptome, particularly stimulating proliferative activity in the endometrium.

Population structuring of the ubiquitous stingless bee Tetragonisca angustula in southern Brazil as revealed by microsatellite and mitochondrial markers

Tetragonisca angustula is one of the most widespread stingless bees in the Neotropics. This species swarms frequently and is extremely successful in urban envi- ronments. In addition, it is one of the most popular stingless bee species for beekeeping in Latin America, so nest transportation and trading is common. Nest transportation can change the genetic structure of the host population, reducing inbreeding and increasing homogenization. Here, we evaluate the genetic structure of 17 geographic populations of T. angustula in southern Brazil to quantify the level of genetic differentiation between populations. Analyses were conducted on partially sequenced mitochondrial genes and 11 microsatellite loci of 1002 workers from 457 sites distributed on the mainland and on 3 islands. Our results show that T. angustula populations are highly differentiated as demon- strated by mitochondrial DNA （mtDNA） and microsatellite markers. Of 73 haplotypes, 67 were population-specific. MtDNA diversity was low in 9 populations but microsatellite diversity was moderate to high in all populations. Microsatellite data suggest 10 genetic clusters and low level of gene flow throughout the studied area. However, physical barri- ers, such as rivers and mountain ranges, or the presence or absence of forest appear to be unrelated to population clusters. Factors such as low dispersal, different ecological con- ditions, and isolation by distance are most likely shaping the population structure of this species. Thus far, nest transportation has not influenced the general population structure in the studied area. However, due to the genetic structure we found, we recommend that nest transportation should only occur within and between populations that are genetically similar.